The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes.
The IS2 Element Improves Transcription Efficiency of Integration-Deficient Lentiviral Vector Episomes.
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DOI：
10.1016/j.omtn.2018.08.007
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发表时间：
2018-12-07
期刊：
影响因子：
--
通讯作者：
Benabdellah K
中科院分区：
文献类型：
--
作者：
Sánchez-Hernández S;Gutierrez-Guerrero A;Martín-Guerra R;Cortijo-Gutierrez M;Tristán-Manzano M;Rodriguez-Perales S;Sanchez L;Garcia-Perez JL;Chato-Astrain J;Fernandez-Valades R;Carrillo-Galvez AB;Anderson P;Montes R;Real PJ;Martin F;Benabdellah K
整合缺陷型慢病毒载体（IDLV）已成为基因治疗应用和基础研究的重要替代工具。不幸的是，IDLV与其整合对应物相比显示出较低的转基因表达。在本研究中，我们的目的是提高表达水平的IDLVs插入IS 2元件，其中含有SAR和HS 4序列，到他们的LTR（SE-IS 2-IDLVs）。与我们的预期相反，IS 2元件的存在并没有消除组蛋白脱乙酰酶的表观遗传沉默。此外，IS 2元件降低了IDLV转导细胞中的附加体水平。有趣的是，尽管有这些负面影响，SE-IS 2-IDLV在几种细胞系（包括神经元、神经元祖细胞和诱导多能干细胞）中转基因的百分比和表达水平方面优于SE-IDLV。我们估计IS 2元件增强IDLV LTR环的转录活性6至7倍。IS 2元件在IDLV中的最终作用将在很大程度上取决于靶细胞以及IS 2元件在每种细胞类型中的负效应与正效应之间的平衡。SE-IS 2-IDLV的更好的性能不是由于1-LTR环与2-LTR环的比例的改善的稳定性或差异，而是可能由于IS 2-附加体重新定位到转录活性区域。
Integration-defective lentiviral vectors (IDLVs) have become an important alternative tool for gene therapy applications and basic research. Unfortunately, IDLVs show lower transgene expression as compared to their integrating counterparts. In this study, we aimed to improve the expression levels of IDLVs by inserting the IS2 element, which harbors SARs and HS4 sequences, into their LTRs (SE-IS2-IDLVs). Contrary to our expectations, the presence of the IS2 element did not abrogate epigenetic silencing by histone deacetylases. In addition, the IS2 element reduced episome levels in IDLV-transduced cells. Interestingly, despite these negative effects, SE-IS2-IDLVs outperformed SE-IDLVs in terms of percentage and expression levels of the transgene in several cell lines, including neurons, neuronal progenitor cells, and induced pluripotent stem cells. We estimated that the IS2 element enhances the transcriptional activity of IDLV LTR circles 6- to 7-fold. The final effect the IS2 element in IDLVs will greatly depend on the target cell and the balance between the negative versus the positive effects of the IS2 element in each cell type. The better performance of SE-IS2-IDLVs was not due to improved stability or differences in the proportions of 1-LTR versus 2-LTR circles but probably to a re-positioning of IS2-episomes into transcriptionally active regions.
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影响因子：
4.2
作者：
Gutierrez-Guerrero, Alejandra;Sanchez-Hernandez, Sabina;Martin, Francisco
通讯作者：
Martin, Francisco
DOI：
10.1038/mtna.2013.47
发表时间：
2013-09-03
期刊：
Molecular therapy. Nucleic acids
影响因子：
--
作者：
通讯作者：
--
影响因子：
3.7
作者：
Arumugam, Paritha I.;Urbinati, Fabrizia;Malik, Punam
通讯作者：
Malik, Punam
影响因子：
3.7
作者：
Dunn, KL;Zhao, H;Davie, JR
通讯作者：
Davie, JR
影响因子：
5.6
作者：
Cai, Shi-Xia;Liu, Ai-Ran;Qiu, Hai-Bo
通讯作者：
Qiu, Hai-Bo